Bioassay Screening of the Ethanolic Extract of Tithonia Diversifolia

INTERNATIONAL JOURNAL OF BIOASSAYS ISSN: 2278-778X CODEN: IJBNHY ORIGINAL RESEARCH ARTICLE OPEN ACCESS Bioassay screening of the ethanolic extract of Tithonia diversifolia leaves on selected microorganisms 1* 2 1 1 Anthoney Swamy T , Jackie Obey K , Miyogo Edwin and Lasiti T. Timothy 1Department of Chemistry, University of Eastern Africa, Baraton, Eldoret, Kenya. 2Departmentof Medical Lab Science, University of Eastern Africa Baraton, Eldoret, Kenya. Received for publication: September 21, 2015; Revised: October 24, 2015; Accepted: October 27, 2015 Abstract: The study was conducted to analyze the antibacterial activity of Tithonia diversifolia leaves. The plant was extracted using Ethanol and water in the ratio 9:1. From the study the plant Tithonia diversifolia was found to inhibit the growth of Staphilococcus epidermidis (36.80mm), Enterobacter aerogenes (31.20mm), Streptococcus α-hemolytic (29.60mm), Bacillus cereus (26.2mm), Escherichia coli (10.40mm) and Streptococcus γ-hemolytic (0.000mm)The data collected and documented in this paper is a scientifically justification that the plant can be used to treat against various diseases caused by Staphilococcus epidermidis and Enterobacter aerogenes. However, further studies need to be done to identify the mode of action of the active compounds in the plant. Key words: Tithonia diversifolia; Antibacterial; Medicinal herbs; Leaves; Ethanol; Aqua. Introduction In continuation with our interest in the study on knowledge have been passed on from generation to medicinal plants (Anthoney et al., 2013; 2014; generation (Adedapo et al., 2005). Even today, a Anthoney et al., 2015; Obey et al., 2014; Obey et al., significant proportion of the populace, particularly 2015), we take up on antibacterial activity of in the developing world depends on herbal ethanolic-aqua extract of Tithonia diversifolia against medicines. This is particularly evident in the rural selected microorganisms. areas where infectious diseases are endemic and modern health care facilities are few and far In an increasing search of new antimicrobial agent between and where the people nurse their ailments to cope with the microbial resistance to antibiotics, back to health using local herbs. scientists are searching from different sources including plants. Plants from different genera and Tithonia diversifolia is significant in phyto-medicine, species were found to have antimicrobial potentials and it is commonly known as Mexican sunflower. which lead to the discovery and development of It is a shrub belonging to the family Asteraceae, new antimicrobials or drugs (Hammer et al., 1999; Order Asterales, genus Tithonia, and species Sharififar et al., 2009; Ilesanmi and Olawoye, 2011). diversifolia with binominal name as Tithonia diversifolia The detection of the antimicrobial properties of a (Hemsl) A. Gray. It is asucculent (scandent) shrub plant indicates that, such plant could be a good 1, 2-3 meters tall, with leaves opposite or alternate. source for the development of antimicrobial agent. Each mature stem may bear several flowers at the From antiquity, nature has been a rich store of top of the branches. The specific name remedies for relief from various ailments affecting “diversifolia” means “separated leaves” from the mankind. Plants, marine organisms and Latin “diversus” (divergent) and folium. microorganisms produce structurally diverse compounds, which are useful as drugs, lead Tithonia originated from Mexico, and it is now structures or raw materials (Adedapo et al., 2005). widely distributed throughout the humid and sub- Plants have been used for thousands of years in humid tropics in Central and South America, Asia traditional medicine. The earliest written records on and Africa (Sonke, 1997), and it is common in Egyptian, Chinese, Indian, Greek and Roman indigenous fallow systems in Southeast Asia (M. traditional medicine have listed medicinal plants Cairns, personal communication). Tithonia was and prescriptions used in treating various ailments. probably introduced into Africa as an ornamental. It has been reported in Kenya (Niang et al., 1996), In Africa, medicinal recipes from plants have been Malawi (Ganunga et al., 1998), Nigeria (Ayeni et al., passed orally from generation to generation 1997), Rwanda (Drechsel and Reck, 1998) and (Adedapo et al., 2005). In resource poor Zimbabwe (Jiri and Waddington, 1998). It is found communities, ignorance to good hygienic practices, in Kenya on road-sides, crop fields and waste areas. poverty coupled with high cost of synthetic drugs and the circulation of drugs of questionable The tradition of plant collection and plant-based qualities and counterfeit pharmaceuticals combine medication has been handed down from generation to worsen the plight of the less privileged, forcing to generation, usually by words of mouth among many to seek for the medicines of their ancestors. many cultures (Gurib-Fakim, 1996). T. diversifolia Herbs have been used as sources of food and has been used in traditional medicine for the medicinal purposes for centuries and this treatment of various ailments (Obafemi, 2006). The *Corresponding Author: Dr. T. Anthoney Swamy, Ph.D Head, Department of Chemistry, University of Eastern Africa, Baraton, P.O. Box 2500, Eldoret, Kenya. 4794 T. Anthoney Swamy et al., International Journal of Bioassays 5.2 (2016): 4794-4798 hot water extract of aerial parts of this plant is used Standardization of microorganisms for the treatment of malaria in Nigeria and other Culture was standardized according to the methods places (Calzada, 1978). An oral decoction of the described by Baker and Thomsberg (1983) and the leaves and stem is used to cure hepatitis in Taiwan National Committee for Clinical Laboratory and gastro-intestinal disorders in Kenya and Standards (NCCLS, 2002). About 0.2 mL of an 18 Thailand (Johns, 1995) Also, the infusion of the hrs old culture of each bacterium was suspended leaves is used for the treatment of measles in into sterile universal bottles containing 20 ml Cameroon (Kamden, 1986). A decoction of the nutrient broth and incubated for 5 hrs at 37°C to flowers is used for the treatment of skin eczema obtain a logarithm growth phase. Normal saline (Gurib-Fakim, 1996). Extracts of the various parts was gradually added so as to compare its turbidity of the plant have been reported to exhibit anti- to McFarland Standard of 0.5 which corresponds malaria (Madureira, 2002), anti-inflammatory to approximately 1.0 × 108 CFU/mL. (Rungeler, 1998), anti-proliferation (Victor, 2004) insecticidal (Mata-Greenwood, 2002) and anti- Preparation of the extract concentration and bacterial (Hongsbhanich, 1999) activities. Some antibiotic researchers (Bork, 1996) reported its cancer growth Stock solutions of the extracts were prepared by inhibitory property. It has been reported also that dissolving 500mg in 1ml of dimethylsulfoxide T. diversifolia improves glucose metabolism by (DMSO). A serial double dilution was prepared for reducing insulin resistance and so may be useful for each extract to obtain 500mg/ml, 250mg/ml, the treatment of Type II diabetes (Miura, 2005). It 125mg/ml, 62.5mg/ml,31.25mg/ml, 15.63mg/ml, remains a fact that much work has been done with 7.81mg/ml, 3.91mg/ml, and 1.95mg/ml the extract from T. diversifolia and a lot of its respectively. An antibiotic control was made by medicinal importance has been over emphasized. dissolving 1µg of positive control in 1ml of sterile In the race to meet the millennium development distilled water. DMSO served as a negative control. goals, combat increasing drug resistance, tackle emergence of new diseases and high cost of Susceptibility testing of bacteria species orthodox medicine, traditional medicine is Susceptibility was determined using the Agar cup successfully making a rapid scientific come back diffusion technique. (Adeniyi et al., 2004) A 0.1 mL globally. aliquot of logarithmic phase broth culture of each bacterium (optical density equivalent to 107-108 The present study was carried out to evaluate the CFU/mL) was used to seed sterile molten Mueller- antibacterial activity of ethanolic extract of Tithonia Hinton agar (Oxoid) medium. The seeded plates diversifolia leaves against selected pathogenic were allowed to dry in the dryer for 20 min. A organisms. standard cork borer (6 mm diameter) was used to cut uniform wells on the surface of the agar, into Materials and Methods which was added increasing concentrations of Extraction procedures reconstituted test extract. A pre-incubation The fresh leaves of Tithonia diversifolia were air-dried diffusion of the extracts into the seeded medium at room temperature and reduced to powdery form was allowed for 1 hr. Bacteria plates were incubated using a laboratory mortar and pestle, a modification at 37°C in an incubator for 18-24 hours after which of Victor et al (Miura, 2002) technique. One diameters of zones of inhibition (mm) were hundred and eighty grams (180 g) of the powdery measured. Since each of the extracts was form was extracted using ethanol and water in the reconstituted in solvents, those diluents were ratio 9:1. The macerated mixture was carefully included in each plate as controls. filtered through What man filter paper size 0.1 micrometer. The filtrate was evaporated in well Results and discussion regulated water bath maintained at 450C, and the Table 1: Zone of inhibition (mm ± S.E.) of dilutions of the extract yielded a green solid extract Ethanolic extract of Tithonia diversifolia leaves Zone of Inhibition Positive Control

Load more